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Estrogen regulation of tissue-specific expression of complement C3
S A Sundstrom1, B S Komm, H Ponce-de-Leon
1Department of Obstetrics and Gynecology, University of Pennsylvania, School of Medicine, Philadelphia 19104.
The Journal of Biological Chemistry
|October 5, 1989
Summary
Estradiol stimulates uterine epithelial cells in rats to produce complement component C3 (C3). This estrogen-induced C3 production is specific to the uterus and not observed in the liver.
Area of Science:
- Endocrinology
- Immunology
- Molecular Biology
Background:
- Estradiol administration to immature rats increases a specific protein synthesis in uterine epithelial cells.
- A 180-kDa protein, composed of 115- and 65-kDa subunits, is secreted by uterine luminal epithelial cells following estradiol treatment.
Purpose of the Study:
- To identify and characterize the estradiol-induced 180-kDa uterine protein.
- To investigate the role of estradiol in regulating gene expression in rat uterine cells.
Main Methods:
- cDNA cloning and sequencing using a monoclonal antibody against the 180-kDa protein.
- Northern hybridization to analyze mRNA levels.
- Immunoprecipitation and Western blotting to confirm protein identity.
- Immunohistochemistry to determine protein localization.
Main Results:
- The cloned cDNA (LE-1) showed high homology to complement component C3 (C3), particularly the alpha subunit.
- Estradiol treatment caused a 25-fold increase in a 6.0-kilobase mRNA in uterine cells, homologous to C3 mRNA.
- Immunoprecipitation confirmed the estradiol-induced secretion of a 180-kDa protein, dissociating into 115- and 65-kDa subunits, identified as C3.
- Immunohistochemistry revealed C3 presence exclusively in estrogen-stimulated uterine epithelial cells.
Conclusions:
- Rat uterine epithelial cells produce complement component C3 (C3) in response to estradiol.
- Estradiol-induced C3 expression is specific to uterine epithelial cells and is not modulated by estrogens in the liver.